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Preparation of chitosan films using different neutralizing solutions to improve endothelial cell compatibility

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Abstract

The development of chitosan-based constructs for application in large-size defects or highly vascularized tissues is still a challenging issue. The poor endothelial cell compatibility of chitosan hinders the colonization of vascular endothelial cells in the chitosan-based constructs, and retards the establishment of a functional microvascular network following implantation. The aim of the present study is to prepare chitosan films with different neutralization methods to improve their endothelial cell compatibility. Chitosan salt films were neutralized with either sodium hydroxide (NaOH) aqueous solution, NaOH ethanol solution, or ethanol solution without NaOH. The physicochemical properties and endothelial cell compatibility of the chitosan films were investigated. Results indicated that neutralization with different solutions affected the surface chemistry, swelling ratio, crystalline conformation, nanotopography, and mechanical properties of the chitosan films. The NaOH ethanol solution-neutralized chitosan film (Chi-NaOH/EtOH film) displayed a nanofiber-dominant surface, while the NaOH aqueous solution-neutralized film (Chi-NaOH/H2O film) and the ethanol solution-neutralized film (Chi-EtOH film) displayed nanoparticle-dominant surfaces. Moreover, the Chi-NaOH/EtOH films exhibited a higher stiffness as compared to the Chi-NaOH/H2O and Chi-EtOH films. Endothelial cell compatibility of the chitosan films was evaluated with a human microvascular endothelial cell line, HMEC-1. Compared with the Chi-NaOH/H2O and Chi-EtOH films, HMECs cultured on the Chi-NaOH/EtOH films fully spread and exhibited significantly higher levels of adhesion and proliferation, with retention of the endothelial phenotype and function. Our findings suggest that the surface nanotopography and mechanical properties contribute to determining the endothelial cell compatibility of chitosan films. The nature of the neutralizing solutions can affect the physicochemical properties and endothelial cell compatibility of chitosan films. Therefore, selection of suitable neutralization methods is highly important for the application of chitosan in tissue engineering.

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Acknowledgments

This study was supported by grants from The National Natural Science Foundation of China (30700848, 30772443), The National Basic Research Program of China (“973 Program”, 2005CB623905) and The National Natural Science Foundation of Beijing (7082090).

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Authors and Affiliations

  1. State Key Laboratory of Biomembrane and Membrane Biotechnology, School of Life Sciences, Tsinghua University, Beijing, 100084, China

    Qing He, Yandao Gong & Xiufang Zhang

  2. Institute of Neurological Disorders, Tsinghua University, Beijing, 100049, China

    Qiang Ao

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  1. Qing He
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Correspondence to Xiufang Zhang.

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He, Q., Ao, Q., Gong, Y. et al. Preparation of chitosan films using different neutralizing solutions to improve endothelial cell compatibility. J Mater Sci: Mater Med 22, 2791–2802 (2011). https://doi.org/10.1007/s10856-011-4444-y

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